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C ell lysis devices are needed

C ell lysis devices are needed. Genetically Engineered organism. Required in many areas One of the most foundational devices. Cell death devices . Odor device Light device Detect device Report device…. Population control New applications. 32,300. 33,200 >>.

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C ell lysis devices are needed

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  1. Cell lysis devices are needed Genetically Engineered organism

  2. Required in many areas One of the most foundational devices Cell death devices Odor device Light device Detect device Report device… • Population control • New applications 32,300 33,200 >>

  3. Whatan unsatisfactorysituation! 27parts Unsatisfied experimental data 6teams To be more Universal !!!

  4. Anti-lysis OFF OFF lysis cassette OFF Expression level Anti-lysisgene lysis cassette Anti-ｌysis >> lysis cassette ON Expression level lysis cassette Anti-lysisgene >>

  5. “Lysisbox”has Two Advantages Opposite function variation Type1: Inducible cell Survive Type2: Inducible cell Death Flexible Threshold

  6. Goals Goals 1Characterizationof Lysis cassette 2Characterization of inducible promoter Lactose promoter Fantastic!? 3Characterization of Anti-Lysis gene Well-characterized constitutive promoter family (BBa_J23xxx)

  7. Goals Goals 1Characterizationof Lysis cassette 2Characterization of inducible promoter Lactose promoter 3Characterization of Anti-Lysis gene Well-characterized constitutive promoter family (BBa_J23xxx)

  8. 1, The lytic system of λphage Lysis cassette Holinmakes a hole in inner membrane Endolysin goes through the hole 3 2 1 1 1 2 2 3 3 Holin Endolysin degrades peptideglycan Endolysin Lysis! Holin Endolysin

  9. 1, Characterization of lysis cassette[method] Lactose promoter Inducer IPTG Holin + Endolysin R0011 Lysis cassette RBS T T Low copy vector Lac repressor over expression

  10. Lysis cassette T T 1, Characterization of lysis cassette[method] Add IPTG Incubate30℃ Incubate30℃ overnight MeasureOD550 Dilute with media

  11. Lysis cassette T T 1, Characterization of lysis cassette[result]

  12. 1.Characterization of lysiscassette [result] １８ｈ

  13. Lysis cassette T T 1, Quantitative characterization [result] 18h OD550

  14. Goals Goals 1Characterizationof Lysis cassette 2Characterization of inducible promoter Lactose promoter lysis⇔ IPTG IPTG⇔RPU lysis⇔ RPU • *RPU(Relative Promoter Units) • iGEM recommends characterizations with RPU • Small Error • Independent of measurement conditions Well -characterized constitutive promoter family (BBa_J23xxx)

  15. 2, Characterization of Plac[R0011] Relative Promoter Unit (RPU)

  16. Lysis cassette T T 2, Quantitative characterization [result] 18h OD550

  17. Lysis cassette T T 2, Quantitative characterization [result] 18h Characterize ability of cell Lysis with RPU!! 0.051 0.106

  18. Goals Goals 1Characterizationof Lysis cassette 2Characterization of inducible promoter Lactose promoter 3Characterization of Anti-Lysis gene Well-characterized constitutive promoter family (BBa_J23xxx)

  19. 3, The System of SΔTMD1 Lysis cassette Anti-killer gene 3 2 1 3 2 1 2 3 1 2 3 Holin Endolysin S SΔTMD1 R

  20. sequences reference SΔTMD1,R,Rz SRRz (lysis cassette) T T 3, Constructing of SΔTMD1 Delition PCR • Deletion PCR • additional experiment

  21. 3.Function check of SΔTMD1 18h TMD1 is essential for lyｔicfunction

  22. 3, characterization of SΔTMD1 Type1: Inducible cell Survive Too weak!! (BBa_J23105) J23105 Neitherworks… Why? Neitherworks… Why? Type2: Inducible cell Death J23105

  23. Goals Goals 1Characterizationof Lysis cassette 2Characterization of inducible promoter Lactose promoter 3Characterization of Anti-Lysis gene Well-characterized constitutive promoter family (BBa_J23xxx)

  24. List Registered parts 18parts submitted

  25. bioremediation aroma biosensor realizes your ideas!!! medicines detergent insecticide art fertilizer

  26. Japan iGEMJapanHuman Practice • attitude survey • on genetic engineering and biotechnology • With • Osaka, KIT-Kyoto, Tokyo-Metropolitan, UT-Tokyo • Total • 1511!!!※Paper-based Q8 Do you know “Synthetic biology”?

  27. Japan iGEMJapanHuman Practice

  28. Gold medal criteria • Characterize or improve an existing BioBrick Part or Device and enter this information back on the Registry. • Help another iGEM team by, for example, characterizing a part, debugging a construct, or modeling or simulating their system. • Develop and document a new technical standard that supports the (i) design of BioBrick Parts or Devices, or (ii) construction of BioBrick Parts or Devices, or (iii) characterization of BioBrick Parts or Devices, or (iv) analysis, modeling, and simulation of BioBrick Parts or Devices, or (v) sharing BioBrick Parts or Devices, either via physical DNA or as information via the internet. • Outline and detail a new approach to an issue of Human Practice in synthetic biology as it relates to your project, such as safety, security, ethics, or ownership, sharing, and innovation.

  29. Acknowledgements Shin-ichiro M. Nomura Shun-ichi Kashida Eri Hayashi Miki Imanishi Ikuhiko Nakase Elizabeth Nakajima Advisors Hirohide Saito Akira Nabetani Yoshihiro Fujita Professor Shin Yonehara Supportors Ken-ichi Yoshikawa Tan Inoue Kunio Takeyasu Masahiro Shirakawa Tokitaka Oyama Miki Ebisuya

  30. Dear Fantastic iGEMers Thankyou!!!

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